Method for preparing vegetable seed active peptide by mixed bacteria solid-state fermentation

A solid-state fermentation and active peptide technology is applied in the field of preparation of rapeseed active peptides, which can solve the problems of destroying the original configuration of amino acids, limited sources of enzymes, severe reaction conditions, etc., and is beneficial to environmental protection and saves biological enzyme preparations. , the effect of simple production process

Inactive Publication Date: 2011-06-01
NANJING UNIV OF FINANCE & ECONOMICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chemical method, that is, when acid or alkali hydrolyzes vegetable protein, chloropropanol carcinogens will be produced, and the reaction conditions are severe, which destroys the original configuration of amino acids. At the same time, the discharge of hydrolysis waste liquid will also cause certain environmental pollution.
Enzymatic hydrolysis is currently the main method for the preparation of rapeseed peptides in my country, but because the anti-nutritional factors in rapeseed meal or rapeseed protein isolate cannot be completely removed, which affects the activity of proteases, the degree of hydrolysis is not high, and the hydrolyzate of rapeseed protein The problem of flavor, especially bitterness, has not been fundamentally resolved. At the same time, the sources of enzymes used for the hydrolysis of rapeseed protein are limited, and the cost remains high. Enzymatic preparation of rapeseed peptides is limited.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Screening of strains for solid-state fermentation of rapeseed meal to produce rapeseed active peptides

[0020] Screening objects: Aspergillus niger, Geotrichum candidum, Aspergillus oryzae, Aspergillus usami, Actinomucor elegans, lactic acid bacteria, Candida utilis, Bacillus licheniformis and Bacillus subtilis.

[0021] Preparation of starter: after the expanded culture of the activated bacterial classification (cultivated by a conventional method), dilute the fermented seeds with sterile water to make the concentration of thalline or spores reach 3×10 7 Each / mL is the starter.

[0022] The double-low rapeseed meal was crushed to a particle size of about 500 μm, and sterilized by moist heat at 121°C for 30 minutes. In the fermenter, the starter is added according to the solid-liquid ratio of 1:1-1:2 (mass ratio), the fermentation temperature is 28°C-35°C, the ambient humidity is 80%-90%, stirring and ventilating. After 3 days of fermentation, distilled wat...

Embodiment 2

[0024] Example 2 Synchronous mixed bacteria solid-state fermentation of rapeseed meal with Bacillus subtilis and Mucor radiata to produce rapeseed peptide

[0025] Preparation of Bacillus subtilis suspension:

[0026] Incline medium: beef extract 3g, peptone 10g, NaCl 5, agar 20g, tap water 1000mL, pH 7.2-7.4.

[0027] Seed medium: beef extract 3g, peptone 10, NaCl 5g, tap water 1000mL, pH 7.2-7.4.

[0028] Nutrient solution composition: glucose 2.6g, KH 2 PO 4 2.6g, sterile water 1000mL, pH 7.0.

[0029] Pick two rings of Bacillus subtilis from the activated strain preservation slant and inoculate it in the seed medium, seal it with 8 layers of gauze, culture it on a shaker at 35°C, 120r / min for 24 hours, and the concentration of bacteria reaches 10 8 individual / mL.

[0030] Dilute the fermented liquid with sterile nutrient solution to prepare the cell concentration of 3×10 7 Individual / mL suspension of Bacillus subtilis.

[0031] Preparation of Mucor actinosa suspensi...

Embodiment 3

[0039] Example 3 Production of rapeseed peptide by solid-state fermentation of rapeseed meal with Mucor radiata and Candida utilis synchronously mixed with bacteria

[0040] Preparation of Mucor actinosa suspension:

[0041] Slant medium: 300g potato, 20g glucose, 20g agar, 1000mL tap water.

[0042]Plate expansion medium: 300g potato, 20g glucose, 20g agar, 1000mL tap water.

[0043] Sterile nutrient solution: glucose 5g, KH 2 PO 4 2g, 1000mL sterile water, pH 6.5.

[0044] Mucor actinosa was inoculated on a slant medium and cultured at a constant temperature of 28°C for 4 days, and then inoculated on a flat plate for expansion culture. After 4 days, the spores were washed with sterile nutrient solution, and the concentration was adjusted to 3×10 7 individual / mL.

[0045] Preparation of Candida utilis suspension:

[0046] Incline medium: 3g of malt extract, 10g of glucose, 3g of yeast extract, 5g of peptone, 20g of agar, 1000mL of tap water.

[0047] Seed medium: malt ...

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Abstract

The invention provides a method for preparing rapeseed active peptide by using mixing bacteria to perform solid state fermentation. The method comprises the following steps: rapeseed pulp obtained by pressing or leaching oil preparation process is crushed to obtain crushed rapeseed pulp; one of bacillus subtilis, lactic acid bacteria or candida utilis is compounded with actinomucor elegans or aspergillus usamii to prepare a fermenting agent; the fermenting agent is injected in the crushed rapeseed pulp for solid state fermentation; a culture medium after solid state fermentation is extracted by adding water, and the residue is removed by separation so as to obtain a coarse extracting solution of the rapeseed active peptide; and the coarse extracting solution is decolored, desalted and dried to obtain the rapeseed active peptide. The method has the advantages of environmental protection and mild reaction conditions, and can fully utilize hydrolytic action and sulfur glycoside degradation action of protease, so the method is low in cost and suitable for industrialized production of the rapeseed active peptide.

Description

technical field [0001] The invention relates to a preparation method of rapeseed active peptide, in particular to a method for preparing rapeseed active peptide through mixed culture of two kinds of microorganisms and solid-state fermentation of rapeseed dregs. Background technique [0002] Rapeseed meal is a by-product of the production of rapeseed oil, which contains 35% to 50% of crude protein, and is an important plant protein resource. Because rapeseed meal contains glucosinolates, erucic acid and other anti-nutritional ingredients, it makes rapeseed cake The application of meal as a vegetable protein resource in the food industry is limited. Rapeseed protein is mainly composed of 12S globulin and 2S or 1.7S albumin. The content of lysine, cystine and methionine is relatively high. The amino acid balance is better than that of soybean protein. It is a high-quality protein. Rapeseed protein is hydrolyzed into polypeptides Finally, it has better biological activity and p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P39/00C12P21/06C07K1/34C07K1/18C12R1/125C12R1/225C12R1/72C12R1/785C12R1/66
Inventor 袁建鞠兴荣何荣王立峰
Owner NANJING UNIV OF FINANCE & ECONOMICS
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